JP4549630B2 - 4- (2-Butylamino) -2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) pyrazolo- [1,5-a] -1 as a corticotropin releasing factor receptor ligand , 3,5-triazine, its enantiomers and pharmaceutically acceptable salts - Google Patents

Description

  The present invention relates to 4- (2-butylamino) -2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) pyrazolo- [1,5-, as corticotropin releasing factor receptor ligands. a] -1,3,5-triazine, its enantiomers and pharmaceutically acceptable salts, to treat major depression, anxiety related disorders, posttraumatic stress disorder, supranuclear palsy and eating disorders It relates to the treatment of psychiatric and neurological disorders, as well as the treatment of immunological, cardiovascular or heart related diseases associated with mental confusion and stress, and colon hypersensitivity.

  Corticotropin-releasing factor (referred to herein as CRF) is a 41 amino acid peptide and is the main physiological regulator of pro-opiomelanocortin (POMC) -derived peptide secretion from the anterior pituitary gland. [(Non-patent document 1); (Non-patent document 2). In addition to its endocrine role in the pituitary gland, hormones have a broad hypothalamic distribution in the central nervous system, and are consistent with the role of neurotransmitters or neuromodulators in the brain. The generation of a broad spectrum of physiological and behavioral effects has been demonstrated by immunohistochemical localization of CRF [(Non-patent document 3; (Non-patent document 4); (Non-patent document 5)]. There is also evidence that CRF plays an important role in integrating the immune system's response to psychological, and immunological stressors [(6); (7)].

  Clinical data provides evidence that CRF has a role in mental and neurological disorders, including depression, anxiety related disorders and eating disorders. Alzheimer's disease, Parkinson's disease, Huntington's disease, progressive supranuclear palsy and amyotrophic lateral sclerosis are associated with dysfunction of CRF neurons in the central nervous system and therefore in their etiology and pathophysiology The role of CRF has been postulated as self-evident [for review see (Non-Patent Document 8)].

  In affective disorder or major depression, the concentration of CRF is significantly increased in the cerebrospinal fluid (CSF) of individuals who do not use drugs [(Non-patent document 9); (Non-patent document 10); Reference 11); ((Non-patent Reference 12)). Furthermore, in response to hypersecretion of CRF, the density of CRF receptors is markedly reduced in the frontal cortex of suicide victims [13]. In addition, in patients with depression, a slowing of the adenocorticotropin (ACTH) response to CRF (intravenous administration) is observed [(Non-patent document 14); (Non-patent document 15); (Non-patent document 16)]. Preclinical studies in rats and non-human primates further supported the hypothesis that hypersecretion of CRF may be responsible for the symptoms seen in human depression [17]. There is preliminary research evidence that tricyclic antidepressants alter CRF levels in the brain and thus regulate the number of CRF receptors [18].

  CRF has also been postulated to have a role in the pathogenesis of anxiety related disorders. It has been demonstrated in various behavioral anxiety models that CRF causes anxiety-inducing action in animals and there is an interaction between benzodiazopine / non-benzodiazepine anxiolytics and CRF [(Non-patent Document 19); (Non-patent document 20)]. Preliminary studies using the putative CRF receptor antagonist a-helical sheep CRF (9-41) in various behavioral paradigms have shown that this antagonist is qualitatively similar to benzodiazepines. "(Non-Patent Document 21), (Non-Patent Document 22)].

  Neurochemical, endocrine, and receptor binding studies all showed an interaction between CRF and benzodiazepine anxiolytics, providing further evidence that CRF is involved in these disorders. Chlordiazepoxide attenuates the “anxiety-inducing” action of CRF in both the rat conflict test [(Non-patent Document 23); (Non-patent Document 24)] and the auditory startle test [(Non-patent Document 25)]. A benzodiazepine receptor antagonist (Ro15-1788), which alone lacked behavioral activity in the operant conflict test, abolished the effects of CRF in a dose-dependent manner, whereas a benzodiazepine inverse agonist (FG7142) did not. [Non-patent document 26]].

  The mechanism and site of action by which standard anxiolytics and antidepressants produce their therapeutic effects must await further elucidation. However, it has been postulated that the drugs are involved in suppressing the hypersecretion of CRF observed in these disorders. Of particular interest is a preliminary study examining the effects of CRF receptor antagonists (a-helical CRF9-41) in various behavioral paradigms, where CRF antagonists are qualitatively similar to benzodiazepines. It has been shown to cause “like” action [for review see (Non-Patent Document 27)].

  In addition, cardiovascular or heart-related diseases and gastrointestinal disorders resulting from stress such as hypertension, tachycardia and congestive heart failure, stroke associated with mental confusion and stress, irritable bowel syndrome, postoperative ileus and colonic hypersensitivity Are assumed to have a role [for review, see (Non-patent Document 28) and (Non-patent Document 29)].

  Overexpression or underexpression of CRF has been proposed as the root cause of several medical diseases. Examples of such treatable disorders include, but are not limited to: affective disorder, anxiety, depression, headache, irritable bowel syndrome, posttraumatic stress disorder, supranuclear palsy, immunosuppression , Alzheimer's disease, gastrointestinal disease, anorexia or other eating disorders, drug addiction, drug or alcohol withdrawal symptoms, inflammatory diseases, cardiovascular or heart related diseases, pregnancy problems, human immunodeficiency virus infection, hemorrhagic Stress, obesity, infertility, head and spinal cord trauma, sputum, stroke, ulcer, amyotrophic lateral sclerosis, hypoglycemia, hypertension, tachycardia and congestive heart failure, mental confusion and stress related stroke, osteoporosis , Premature birth, psychosocial dwarfism, stress-induced fever, ulcer, diarrhea, postoperative ileus and colonic hypersensitivity [For review, (Non-patent document 30), (Non-patent document 3) ), (Non-patent Document 32), (Non-patent Document 33), (Non-patent Document 34), (Non-patent Document 35), see (Non-Patent Document 36) and (Non-Patent Document 37).

  The following publications describe CRF antagonist compounds, but none disclose the compounds described herein: (Patent Literature 1), (Patent Literature 2), (Patent Literature 3), (Patent Literature 4), (Patent Literature 5), (Patent Literature 6), (Patent Literature 7), (Patent Literature 8), (Patent Literature 9), (Patent Literature 10) and (Patent Literature 11).

WO 95/10506 specification WO 99/51608 Specification WO 97/35539 specification WO 99/01439 Specification WO 97/44308 specification WO 97/35846 specification WO 98/03510 specification WO 99/11643 specification PCT / US99 / 18707 specification WO 99/01454 Specification WO 00/01675 Specification J. et al. Rivier et al., Proc. Nat. Acad. Sci. (USA) 80: 4851 (1983) W. Vale et al., Science 213: 1394 (1981) W. Vale et al., Rec. Prog. Horm. Res. 39: 245 (1983) G. F. Koob, Persp. Behav. Med. 2:39 (1985) E. B. De Souza et al. Neurosci. 5: 3189 (1985) J. et al. E. Blackock, Physiological Reviews 69: 1 (1989) J. et al. E. Morley, Life Sci. 41: 527 (1987) E. B. De Souza, Hosp. Practice 23:59 (1988) C. B. Nemerov et al., Science 226: 1342 (1984). C. M.M. Banki et al., Am. J. et al. Psychiatry 144: 873 (1987) R. D. France et al., Biol. Psychiatry 28:86 (1988) M.M. Arato et al., Biol Psychiatry 25: 355 (1989) C. B. Nemerov et al., Arch. Gen. Psychiatry 45: 577 (1988) P. W. Gold et al., Am J. et al. Psychiatry 141: 619 (1984) F. Holsboer et al., Psychoneuroendocrinology 9: 147 (1984). P. W. Gold et al., New Eng. J. et al. Med. 314: 1129 (1986) R. M.M. Sapolsky, Arch. Gen. Psychiatry 46: 1047 (1989) Grigoriadis et al., Neurosychopharmacology 2:53 (1989). D. R. Britton et al., Life Sci. 31: 363 (1982) C. W. Berridge and A.M. J. et al. Dunn Regul. Peptides 16:83 (1986) C. W. Berridge and A.M. J. et al. Dunn Horm. Behav. 21: 393 (1987) Brain Research Reviews 15:71 (1990) K. T.A. Britton et al., Psychopharmacology 86: 170 (1985). K. T.A. Britton et al., Psychopharmacology 94: 306 (1988). N. R. Swedlow et al., Psychopharmacology 88: 147 (1986). K. T.A. Britton et al., Psychopharmacology 94: 306 (1988). G. F. Kobb and K.K. T.A. Britton, Corticotropin-Release Factor: Basic and Clinical Studies of a Neuropeptide, E.M. B. De Souza and C.I. B. Edited by Nemerov, CRC Press 221 (1990) Corticotropin-Release Factor: Basic and Clinical Studies of a Neuropeptide, E.C. B. De Souza and C.I. B. Nemerov, CRC Press p221 (1990) C. Maillot, M.M. Million, J.M. Y. Wei, A .; Gauthier, Y. et al. Tache, Gastroenterology, 119, 1569-1579 (2000). J. et al. R. McCarthy, S.M. C. Heinrichs and D.H. E. Grigoriadis, Cuur. Pharm. Res. , 5, 289-315 (1999) P. J. et al. Gilligan, D.C. W. Robertson and R.C. Zaczek, J. et al. Medicinal Chem. , 43, 1641-1660 (2000) G. P. Crousos, Int. J. et al. Obesity, 24, Suppl. 2, S50-S55 (2000) E. Webster, D.C. J. et al. Torpy, I.D. J. et al. Elenkov, G.M. P. Crousos, Ann. N. Y. Acad. Sci. , 840, 21-32 (1998) D. J. et al. Newport and C.I. B. Nemerov, Curr. Opin. Neurobiology, 10, 211-218 (2000) G. Mastorakos and I. Ilias, Ann. N. Y. Acad. Sci. , 900, 95-106 (2000) M.M. J. et al. Owens and C.I. B. Nemerov, Expert Opin. Invest. Drugs, 8, 1849-1858 (1999) G. F. Koob, Ann. N. Y. Acad. Sci. , 909, 170-185 (2000) Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, PA, 1985, p. 1418 Design of prodrugs, H.M. Bundgaard, Hen, Elsevier, 1985 Methods in Enzymology, K.M. Wider et al., Academic Press, 42, 309-396, 1985. A Textbook of Drug Design and Development, Krogsgaard-Larsen and H.M. Bundgaard ed., Chapter 5; “Design and Applications of prodrugs”, pages 113-191, 1991. Advanced Drug Delivery Reviews, H.C. Bundgard, 8, pp. 1-38, 1992 Journal of Pharmaceutical Sciences, 77, 285, 1988. Chem. Pharm. Bull. , N.M. Nakaya et al., 32, 692, 1984; prodrugs as Novel Delivery Systems, T .; Higuchi and V. Stella, A .; C. S. Volume 14 of Symposium Series Bioreversible Carriers in Drug Design, Edward B. et al. Edited by Roche, American Pharmaceutical Association and Pergamon Press, 1987 Kuhar, Current Protocols in Pharmacology (1998) John Wiley & Sons, New York, Sections 8.1.1-8.1.9. D. Perrin and W. L. F. Armarego, Purification of Laboratory Chemicals, 3rd edition (New York: Pergamon Press, 1988) E. B. De Souza, J .; Neuroscience, 7:88 (1987) P. J. et al. Munson and D.M. Rodbard, Anal. Biochem. 107: 220 (1980) G. Battaglia et al., Synapse 1: 572 (1987) C. W. Berridge and A.M. J. et al. Dunn, Brain Research Reviews 15:71 (1990) Remington's Pharmaceutical Sciences, A.M. Osol

  According to one aspect, the invention relates to affective disorders, anxiety, depression, irritable bowel syndrome, post-traumatic stress disorder, supranuclear palsy, immunosuppression, Alzheimer's disease, gastrointestinal disease, or other eating disorders, Disorders including, but not limited to, drugs or alcohol withdrawal symptoms, drug addiction, inflammatory diseases, pregnancy problems, disorders induced by CRF or promoted by CRF The above disorders, or disorders selected from inflammatory diseases such as rheumatoid arthritis and osteoarthritis, disorders selected from pain, asthma, psoriasis and allergies; generalized anxiety disorder; panic, fear Post-traumatic stress disorder; stress-induced sleep disorder; pain perception such as fibromyalgia; major depression, single episode depression, recurrent depression Mood disorders such as depression, including child abuse-induced depression, and postpartum depression; mood modulation; bipolar disorder; mood circulation; fatigue syndrome; stress-induced headache; cancer, human immunodeficiency virus (HIV) infection; Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and Huntington's disease; ulcers, irritable bowel syndrome, Crohn's disease, convulsive colon, diarrhea, and postoperative ileus and colonic hypersensitivity associated with mental confusion or stress Gastrointestinal diseases such as anorexia nervosa and bulimia nervosa; hemorrhagic stress; stress-induced psychotic episodes; pseudohypothyroid syndrome; due to inappropriate antihormonal hormone (ADH) Symptoms; obesity; infertility; head injury; spinal cord injury; ischemic nerve injury (eg, cerebral hippocampal ischemia) Cerebral ischemia, etc.); excitotoxic nerve injury; epilepsy; cardiovascular and heart related disorders including hypertension, tachycardia and congestive heart failure; stroke; stress-induced immune dysfunction (eg, stress-induced fever, porcine stress Immune dysfunction including syndrome, bovine fever, equine paroxysmal fibrillation, and restraint-induced dysfunction in chickens, hair-hunting stress in sheep or human-animal interaction-related stress in dogs; muscle spasm; urinary incontinence; Alzheimer-type senile dementia; cerebrovascular dementia; amyotrophic lateral sclerosis; drug addiction and drug addiction (eg, dependence on alcohol, cocaine, heroin, benzodiazepines, or other drugs); drugs and alcohol Used in the treatment of withdrawal symptoms; osteoporosis; psychosocial dwarfism; and hypoglycemia in mammals It can be found novel compounds, to provide a pharmaceutical composition and method.

  The present invention provides novel compounds that bind to the corticotropin releasing factor receptor, thereby altering the anxiety-inducing action of CRF secretion. The compounds of the present invention treat psychiatric and neurological disorders, anxiety-related disorders, post-traumatic stress disorder, supranuclear palsy and eating disorders, and immunologically related to mental disruption and stress in mammals, Useful in the treatment of cardiovascular or heart related diseases and colonic hypersensitivity.

  According to another aspect, the present invention provides novel compounds of formula (I) (described below) that are useful as antagonists of corticotropin releasing factor. The compound of the present invention exhibits activity as a corticotropin releasing factor antagonist and is thought to suppress excessive CRF secretion. The present invention also includes pharmaceutical compositions containing such compounds of formula (I), and methods of using such compounds for the suppression of hypersecretion of CRF and / or the treatment of anxiety disorders.

  The use of a competitive binding assay has a greater or more selective binding affinity for a new drug candidate, eg, for a new CRF ligand, or CRF receptor, so the candidate is It is believed to be particularly useful in identifying other compounds that are potentially useful as drugs. In this assay, the ability of a candidate ligand to replace a labeled compound is determined.

  Thus, another embodiment of the invention is a binding assay in which one or more of the compounds are connected to a label, which can provide a detectable signal directly or indirectly. Including the use of compounds. Various labels include radioisotopes, fluorescent materials, chemiluminescent materials, specific binding molecules, particles such as magnetic particles, and the like.

  Another embodiment of the present invention is a standard and reagent for use as a probe to localize receptors in cells and tissues, and for determining the receptor-binding properties of test compounds. It relates to the use of a compound according to the invention, in particular a labeled compound according to the invention. The labeled compounds of the present invention can be used for in vitro tests such as autoradiography of tissue sections or for in vivo methods such as PET or SPECT scanning. In particular, preferred compounds of the invention are useful as standards and reagents in determining the ability of potential pharmaceutical agents to bind to the CRF1 receptor.

およびその立体異性体形、またはその立体異性体形の混合物、およびその薬学的に許容できる塩またはプロドラッグ形を提供する。 [1] In a first embodiment, the present invention provides a compound of formula (I):

And stereoisomeric forms thereof, or a mixture of stereoisomeric forms thereof, and pharmaceutically acceptable salt or prodrug forms thereof.

  [2] In another embodiment, the invention provides that the above compound is 4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3,5-triazine or 4-((S) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl ) [1,5-a] -pyrazolo-1,3,5-triazine, the compound of embodiment [1], its isomer, its stereoisomer form, a mixture of its stereoisomer forms, its pharmaceutically acceptable Prodrugs, or pharmaceutically acceptable salt forms thereof, are provided.

  [3] In another embodiment, the present invention provides the compound according to any one of Embodiments [1] to [2], wherein the compound is substantially free of (S) stereoisomer, Provided are acceptable prodrugs, or pharmaceutically acceptable salt forms thereof.

  [4] In another embodiment, the present invention provides that the above compound is 4- (2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5 -A]-The compound of embodiment [1], which is pyrazolo-1,3,5-triazine.

  [5] In another embodiment, the invention provides that the above compound is 4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) A compound of embodiment [1] is provided that is [1,5-a] -pyrazolo-1,3,5-triazine.

  [6] A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of any one compound of the embodiments [1] to [5].

  [7] In another embodiment, the present invention provides a CRF receptor in a mammal comprising administering to the mammal a therapeutically effective amount of any one of the embodiments [1] to [5]. A method of antagonizing is provided.

  [8] In another embodiment, the present invention relates to hypersecretion of CRF in a warm-blooded animal, comprising administering to the animal a therapeutically effective amount of any one compound of embodiments [1] to [5]. A method of treating a disorder presenting is provided.

  [9] In another embodiment, the present invention is a method of treating a disorder comprising administering to a mammal a therapeutically effective amount of any one compound of embodiments [1] to [5]. Thus, providing a method wherein treatment of the disorder can be effected or promoted by antagonizing CRF.

  [10] In another embodiment, the invention provides a CRF receptor in a mammal comprising administering to the mammal a therapeutically effective amount of any one of the embodiments [1] to [5]. A method of antagonizing is provided.

  [11] In another embodiment, the present invention provides anxiety or depression in a mammal, comprising administering to the mammal a therapeutically effective amount of any one compound of embodiments [1] to [5]. Provide a method of treatment.

[12] In another embodiment, the present invention provides a method for screening a ligand for a CRF receptor comprising:
a) performing a competitive binding assay using the CRF receptor, the compound of any one of embodiments [1]-[5] labeled with a detectable label, and the candidate ligand;
b) determining the ability of the candidate ligand to replace the labeled compound;
Providing a method.

[13] In another embodiment, the present invention provides:
a) contacting one of the compounds of embodiments [1] to [5] labeled with a detectable label under conditions that allow the compound to bind to the tissue;
b) providing a method of detecting a CRF receptor in a tissue comprising detecting a labeled compound bound to the tissue.

  [14] In another embodiment, the invention includes contacting a compound of any one of embodiments [1] to [5] with a solution comprising cells that express the CRF1 receptor. A method of inhibiting binding to CRF-1 receptor, wherein said compound is present in said solution at a concentration sufficient to inhibit binding of CRF to CRF-1 receptor To do.

[15] In another embodiment, the present invention provides:
a) packaging materials;
b) any one of the embodiments [1] to [5];
c) a label, or a package insert included within the packaging material indicating that the compound is effective in treating anxiety or depression;
Providing products, including

  [16] The present invention relates to affective disorder, anxiety, depression, headache, irritable bowel syndrome, comprising administering to a mammal a therapeutically effective amount of any one compound of Embodiments [1] to [5] Post-traumatic stress disorder, supranuclear palsy, immunosuppression, Alzheimer's disease, gastrointestinal disease, anorexia or other eating disorders, drug addiction, drug or alcohol withdrawal symptoms, inflammatory diseases, cardiovascular or heart related diseases, Problems related to pregnancy, human immunodeficiency virus infection, hemorrhagic stress, obesity, infertility, head and spinal cord trauma, epilepsy, stroke, ulcer, amyotrophic lateral sclerosis, hypoglycemia, or induced by CRF A disorder, including, but not limited to, a disorder wherein the treatment of the disorder can be effected or promoted by antagonizing CRF in a mammal, A method of treating also be included.

Definitions As used herein, the term “pharmaceutically acceptable salts” refers to salts prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Suitable non-toxic acids include organic acids with basic residues such as inorganic and amines such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethenesulfonic acid, fumaric acid, gluconic acid, Glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucinic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p- And organic salts of acidic residues such as alkali or carboxylic acid, for example, alkali and alkaline earth metal salts derived from the following bases: sodium hydride, sodium hydroxide, potassium hydroxide, water Calcium oxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, ammonia, Limethylammonia, triethylammonia, ethylenediamine, n-methyl-glucamine, arginine, ornithine, choline, N, N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, n-benzylphenethylamine, diethylamine, piperazine, tris (hydroxymethyl) ) -Aminomethane, tetramethylammonium hydroxide, and the like.

  The pharmaceutically acceptable salts of the compounds of the present invention may be prepared by converting the free acid or base form of these compounds, in water, in an organic solvent, or in a mixture of the two, to a suitable base or acid stoichiometry. In general; non-aqueous medium-like ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. (Non-Patent Document 38) (the disclosure of which is incorporated herein by reference) provides a list of suitable salts.

  As used herein, a “pharmaceutically acceptable prodrug” is a covalent that releases an active parent drug of formula (I) in vivo when such prodrug is administered to a mammalian subject. Means bound carrier. Prodrugs of compounds of formula (I) are within the scope of sound medical evaluation and are suitable for use in contact with human and lower animal tissues with excessive toxicity, irritation, allergic reactions, etc. Suitable for moderate benefit / risk ratios, effective for their intended use and, where possible, the zwitterionic form of the compounds of the invention. The term “prodrug” means a compound that is readily converted in vivo to yield the parent compound of formula (I), for example by hydrolysis in blood. Functional groups that can be readily converted by metabolic cleavage form groups of types that react well with the carboxyl groups of the compounds of the invention in vivo. They are alkanoyl (eg acetyl, propionyl, butyryl etc.), unsubstituted and substituted aroyl (eg benzoyl and substituted benzoyl), alkoxycarbonyl (eg ethoxycarbonyl), trialkylsilyl (eg trimethylsilyl and triethylsilyl) ), Monoesters formed by dicarboxylic acids (eg, succinyl), and the like, but are not limited thereto. Since the metabolically cleavable groups of the compounds useful according to the present invention are readily cleaved in vivo, the compounds having such groups serve as prodrugs. The present compounds having metabolically cleavable groups exhibit improved bioavailability as a result of the high solubility and / or absorption rate afforded to the parent compound by the presence of metabolically cleavable groups. It has the advantage of being possible. In the following, prodrugs are fully discussed: (Non-patent document 39), (Non-patent document 40), (Non-patent document 41), (Non-patent document 42), (Non-patent document 43), ( Non-Patent Document 44), and (Non-Patent Document 45), which are hereby incorporated by reference.

  A “prodrug” is considered a covalently bonded carrier that releases the active parent drug of formula (I) in vivo when such prodrug is administered to a mammalian subject. Prodrugs of compounds of formula (I) are prepared by modifying functional groups present in the compound, either routinely or in vivo, in such a way that the modification is cleaved to the parent compound. Is done. A prodrug is a compound in which a hydroxy group, an amine group, or a sulfhydryl group is bonded to a group that cleaves to form a free hydroxyl group, amino group, or sulfhydryl group, respectively, when administered to a mammalian subject. including. Examples of prodrugs include, but are not limited to, acetic acid derivatives, formic acid derivatives, benzoic acid derivatives, and the like of alcohol and amine functional groups in compounds of formula (I).

  The term “substantially free of its (S) stereoisomer” as used herein to describe a compound refers to the (S) stereoisomer, where the compound is an optical enantiomer. It means that it is composed of the (R) stereoisomer more significantly. In a preferred embodiment of the invention, the term “substantially free of its (S) stereoisomer” means that the compound is at least about 90% by weight of its (R) stereoisomer, and about 10% by weight. It means to be composed of the following (S) stereoisomers.

  In a more preferred embodiment of the invention, the term “substantially free of its (S) stereoisomer” means that the compound is at least about 95% by weight of its (R) stereoisomer, and about 5 It means that it is composed of the (S) stereoisomer in an amount of not more than% by weight. In an even more preferred embodiment, the term “substantially free of its (S) stereoisomer” means that the compound is at least about 99% by weight of its (R) stereoisomer, and no more than about 1%. , (S) means that it is composed of stereoisomers. In another preferred embodiment, the term “substantially free of its (S) stereoisomer” means that the compound is composed of approximately 100% by weight of its (R) stereoisomer. . The above percentages are based on the total amount of combined stereoisomers of the compound.

  The term “therapeutically effective amount” of a compound of the invention means an effective amount to antagonize abnormal levels of CRF or to treat symptoms of host affective disorder, anxiety or depression.

As used herein, the term “labeled” refers to a label by which the compound provides a detectable signal, eg, particles such as radioisotopes, fluorescent materials, enzymes, antibodies, magnetic particles, chemiluminescent materials, etc. , P ³² , I ¹³¹ , At ^{211 and the} like.

Synthetic Many organic compounds exist in optically active forms, ie have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule around its chiral center. The prefixes d and l or (+) and (−) are used to indicate the sign of rotation of plane polarized light by the compound, (−) or l means that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these compounds, called stereoisomers, are identical except that they are mirror images of one another. Certain stereoisomers are also referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture.

  The present invention includes all stereoisomeric forms of the compounds of formula I. All asymmetric centers present in compounds of formula I have the S or R configuration, independent of one another. The prefixes d and l or (+) and (−) are used to indicate the sign of rotation of plane polarized light by the compound, (−) or l means that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. The present invention includes all possible enantiomers and diastereomers, and mixtures of two or more stereoisomers, eg, mixtures of enantiomers and / or diastereomers in any ratio. Thus, enantiomers, both levorotatory and dextrorotatory enantiomers, are in enantiomerically pure form, in racemic form, and in the form of a mixture of two enantiomers in any ratio. This is the subject of the present invention. In the case of cis / trans isomerism, the present invention includes both cis and trans forms, and mixtures of these forms in any ratio. If necessary, the individual stereoisomers can be synthesized by customary methods such as resolution of the mixture by chromatography or crystallization, by using stereochemically homogeneous starting materials in the synthesis, or by stereoselective synthesis. Can be prepared. Optionally, derivatization may be performed prior to resolution of the stereoisomers. Resolution of the mixture of stereoisomers can be carried out at the stage of the compound of formula I or at the intermediate stage during synthesis. The present invention also includes all tautomeric forms of the compounds of formula (I).

Compounds of formula (I) can be prepared using the procedure outlined in Scheme 1.

  In an inert solvent, a compound of formula (II) where X = F is added with a metal alkoxide (eg sodium methoxide, potassium methoxide; preformed or generated in situ) to give a compound of formula (III) The intermediate of Inert solvents include alkyl alcohols (1-8 carbons, preferably methanol or ethanol), lower alkanenitriles (1-6 carbons, preferably acetonitrile), water, dialkyl ethers (preferably Diethyl ether), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), cyclic amides (Preferably N-methylpyrrolidin-2-one), dialkyl sulfoxides (preferably dimethyl sulfoxide) or aromatic hydrocarbons (preferably benzene or toluene) are included, but not limited thereto. A preferred reaction temperature is in the range of 0 ° C to 100 ° C.

Alternatively, an alkylating agent may be added to a compound of formula (II) where X = OH in the presence of a base in an inert solvent to give an intermediate of formula (III). Alkylating agents include, but are not limited to, haloalkanes (eg CH ₃ I), dialkyl sulfates (eg Me ₂ SO ₄ ) or alkyl trifluoro-sulfonates (eg CH ₃ O ₃ SCF ₃ ). .

  Bases include alkali metals, alkali metal hydrides (preferably sodium hydride), alkali metal alkoxides (1-6 carbons) (preferably sodium methoxide or sodium ethoxide), alkaline earth metal hydrides, Alkali metal carbonates, alkaline earth metal carbonates, transition metal carbonates (eg silver carbonate), alkali metal dialkylamides (preferably lithium diisopropylamide), alkali metal bicarbonates, alkali metal hydroxides, alkali metal bis (Trialkylsilyl) amides (preferably sodium bis (trimethylsilyl) amide), trialkylamines (preferably N, N-di-isopropyl-N-ethylamine) or aromatic amines (preferably pyridine) But not limited to

  Inert solvents include halocarbons (1-8 carbons, 1-8 halogens), lower alkanenitriles (1-6 carbons, preferably acetonitrile), water, dialkyl ethers (preferably Is diethyl ether), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), cyclic amides (Preferably N-methylpyrrolidin-2-one), dialkyl sulfoxides (preferably dimethyl sulfoxide), aromatic hydrocarbons (preferably benzene or toluene) and the like. The preferred reaction temperature is in the range of 50 ° C to 150 ° C.

A compound of formula (III) can be converted to a compound of formula (IV) by reaction with a brominating agent in an inert solvent in the presence or absence of an additive. Examples of brominating agents include N-bromosuccinimide-2,2'-azobisisobutyronitrile (AIBN), N-bromophthalimide-2,2'-azobisiso-butyronitrile (AIBN)), bromine and the like. However, it is not limited to these. Examples of additives include alkali metal phosphates (for example, K ₃ PO ₄ , Na ₃ PO ₄ ), alkali metal hydrogen phosphates (for example, Na ₂ HPO ₄ , K ₂ HPO ₄ ), alkali metal dihydrogen salts ( Examples thereof include, but are not limited to, NaH ₂ PO ₄ , KH ₂ PO ₄ ). Inert solvents include halocarbons (1-6 carbons, 1-6 halogens (preferably chlorine)), water, N, N-dialkylformamides (preferably dimethylformamide), N, N -Dialkylacetamides (preferably dimethylacetamide), cyclic amides (preferably N-methylpyrrolidin-2-one) and the like are included, but are not limited thereto. The reaction temperature is in the range of 0 ° C to 200 ° C (preferably 20 ° C to 120 ° C).

The compound of formula (IV) is (1) alkyllithium in an inert solvent at a temperature in the range of −100 ° C. to 50 ° C. and (2) at a temperature in the range of −100 ° C. to 50 ° C. (OR ^a ) ₃ (R ^a is a branched or straight chain alkyl having 1 to 20 carbons) and (3) -100 ° C to 100 ° C in the presence or absence of water. It can be converted to the compound of formula (V) by successive reaction with acid at a temperature in the range. Alkyllithium may be a branched or linear compound containing 1 to 20 carbons. Examples of the inert solvent include dialkyl ethers (preferably diethyl ether), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), or aromatic hydrocarbons (preferably benzene or toluene). It is not limited to these.

  Acids include alkanoic acids having 2 to 10 carbons (preferably acetic acid), haloalkanoic acids (2 to 10 carbons, 1 to 10 halogens such as trifluoroacetic acid), aryl sulfonic acids (preferably p-toluenesulfonic acid or benzenesulfonic acid), alkanesulfonic acid having 1 to 10 carbons (preferably methanesulfonic acid), hydrochloric acid, sulfuric acid, phosphoric acid, and the like are included, but are not limited thereto.

A compound of formula (VII) can be prepared by reacting a compound of formula (V) with a compound of formula (VI) in the presence of a palladium or nickel complex or salt, a base and an inert solvent. it can. Palladium or nickel complexes include Pd (PPh ₃ ) ₄ , PdCl ₂ (PPh ₃ ) ₂ , NiCl ₂ (PPh ₃ ) ₂ , or [1,1-bis (diphenylphosphino) ferrocene] -dichloropalladium Including, but not limited to, phosphine complexes. Bases include alkali metals, alkali metal hydrides (preferably sodium hydride), alkali metal alkoxides (1-6 carbons) (preferably sodium methoxide or sodium ethoxide), alkali metal carbonates, alkaline earths. Metal carbonates (such as barium carbonate), transition metal carbonates (such as silver carbonate), or trialkylamines (such as triethylamine), but are not limited thereto. Examples of the inert solvent include dialkyl ethers (preferably diethyl ether), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), or aromatic hydrocarbons (preferably benzene or toluene). It is not limited to these. The preferred reaction temperature is in the range of -100 ° C to 100 ° C.

  The intermediate of formula (VII) can be reacted with a base in the presence of an inert solvent to give a compound of formula (VIII) where M is an alkali metal cation (eg sodium or potassium). is there. Bases include alkali metal hydroxides (eg, NaOH or KOH), alkali metal alkoxides (1-6 carbons) (preferably sodium methoxide or sodium ethoxide), alkaline earth metal hydroxides, etc. However, it is not limited to these. Inert solvents include alkyl alcohols (1-6 carbons), lower alkanenitriles (1-6 carbons, preferably acetonitrile), water, cyclic ethers (preferably tetrahydrofuran or 1,4-carbon. Dioxane), N, N dialkylformamides (preferably dimethylformamide), N, N dialkylacetamides (preferably dimethylacetamide), cyclic amides (preferably N-methylpyrrolidin-2-one), dialkyl sulfoxides (preferably Includes dimethyl sulfoxide), but is not limited thereto. The preferred reaction temperature is in the range of 0 ° C to 150 ° C.

  The compound of formula (VIII) is treated with hydrazine hydrate in the presence of an acid and an inert solvent at a temperature in the range of 0 ° C. to 200 ° C., preferably 70 ° C. to 150 ° C., to give a compound of formula (IX) It is possible to produce compounds. Acids include alkanoic acids having 2 to 10 carbons (preferably acetic acid), haloalkanoic acids (2 to 10 carbons, 1 to 10 halogens such as trifluoroacetic acid), aryl sulfonic acids (preferably p -Toluenesulfonic acid or benzenesulfonic acid), including but not limited to alkanesulfonic acids having 1 to 10 carbons (preferably methanesulfonic acid), hydrochloric acid, sulfuric acid or phosphoric acid.

  Inert solvents include water, alkyl alcohols (1-8 carbons, preferably methanol or ethanol), lower alkanenitriles (1-6 carbons, preferably acetonitrile), cyclic ethers (preferably Tetrahydrofuran or 1,4-dioxane), N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), cyclic amides (preferably N-methylpyrrolidine-2- ON), dialkyl sulfoxides (preferably dimethyl sulfoxide) or aromatic hydrocarbons (preferably benzene or toluene), and the like.

A compound of formula (IX) is prepared in the presence of an acid in the presence or absence of an acid at a temperature ranging from 0 ° C. to 200 ° C. with the formula H ₃ C (C═NH) OR ^C (R ^c is Can be reacted with a compound of alkyl (1-6 carbons) to produce a compound of formula (X). Acids include alkanoic acids having 2 to 10 carbons (preferably acetic acid), haloalkanoic acids (2 to 10 carbons, 1 to 10 halogens such as trifluoroacetic acid), aryl sulfonic acids (preferably p-toluenesulfonic acid or benzenesulfonic acid), alkanesulfonic acid having 1 to 10 carbons (preferably methanesulfonic acid), hydrochloric acid, sulfuric acid, phosphoric acid, and the like are included, but are not limited thereto. It is possible to use stoichiometric or catalytic amounts of such acids.

  Inert solvents include water, alkanenitriles (1-6 carbons, preferably acetonitrile), halocarbons having 1-6 carbons and 1-6 halogens (preferably dichloroethane or chloroform). Alkyl alcohols having 1 to 10 carbons (preferably ethanol), dialkyl ethers (4 to 12 carbons, preferably diethyl ether or di-isopropyl ether) or cyclic ethers such as dioxane or tetrahydrofuran, etc. Including, but not limited to. A preferred temperature is in the range of 0 ° C to 100 ° C.

The compound of formula (X) is compound C═O (R ^d ) ₂ (R ^d is R ^d ) in an inert solvent, in the presence or absence of a base, at a reaction temperature ranging from −50 ° C. to 200 ° C. , Halogen (preferably chlorine), alkoxy (1-4 carbons) or alkylthio (1-4 carbons) can be converted to an intermediate compound of formula (XI) It is. Bases include alkali metal hydrides (preferably sodium hydride), alkali metal alkoxides (1-6 carbons) (preferably sodium methoxide or sodium ethoxide), alkali metal carbonates, alkali metal hydroxides. , Trialkylamines (preferably N, N-di-isopropyl-N-ethylamine or triethylamine) or aromatic amines (preferably pyridine).

  Inert solvents include alkyl alcohols (1-8 carbons, preferably methanol or ethanol), lower alkanenitriles (1-6 carbons, preferably acetonitrile), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), cyclic amides (preferably N-methylpyrrolidin-2-one) , Dialkyl sulfoxides (preferably dimethyl sulfoxide) or aromatic hydrocarbons (preferably benzene or toluene), and the like.

The compound of formula (XI) is treated with a halogenating agent and in the presence or absence of a base, in the presence or absence of an inert solvent, at a reaction temperature ranging from −80 ° C. to 250 ° C. Halogenated intermediate (XII) (X is a halogen) can be obtained. As the halogenating _{agent, SOCl 2, POCl 3, PCl} 3, PCl 5, POBr 3, but it is like PBr ₃ or PBr _5, not limited thereto. Bases include, but are not limited to, trialkylamines (preferably N, N-di-isopropyl-N-ethylamine or triethylamine) or aromatic amines (preferably N, N-diethylaniline). .

  Examples of the inert solvent include N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), and cyclic amides (preferably N-methylpyrrolidin-2-one). Alternatively, aromatic hydrocarbons (preferably benzene or toluene) are included, but not limited thereto. A preferred reaction temperature is a temperature in the range of 20 ° C to 200 ° C.

  A compound of formula (XII) is reacted with an alkylamine in the presence or absence of a base, in the presence or absence of an inert solvent, at a reaction temperature in the range of −80 to 250 ° C. It is possible to give compounds of (I). Bases include alkali metal hydrides (preferably sodium hydride), alkali metal alkoxides (1 to 6 carbons) (preferably sodium methoxide or sodium ethoxide), alkaline earth metal hydrides, alkali metal dialkyls. Amides (preferably lithium di-isopropylamide), alkali metal carbonates, alkali metal bicarbonates, alkali metal bis (trialkylsilyl) amides (preferably sodium bis (trimethylsilyl) amide), trialkylamines (preferably Include, but are not limited to, N, N-di-isopropyl-N-ethylamine) or aromatic amines (preferably pyridine).

  Inert solvents include alkyl alcohols (1-8 carbons, preferably methanol or ethanol), lower alkanenitriles (1-6 carbons, preferably acetonitrile), dialkyl ethers (preferably diethyl ether). ), Cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), N, N-dialkylformamides (preferably dimethylformamide), N, N-dialkylacetamides (preferably dimethylacetamide), cyclic amides (preferably N-methylpyrrolidin-2-one), dialkyl sulfoxides (preferably dimethyl sulfoxide), aromatic hydrocarbons (preferably benzene or toluene) or haloalkanes having 1 to 10 carbons and 1 to 10 halogens Preferably including but dichloroethane), and the like. The preferred reaction temperature is in the range of 0 ° C to 140 ° C.

The compounds of the present invention can be prepared as radiolabeled compounds by performing their synthesis using precursors containing at least one atom that is a radioisotope. The radioisotope is preferably selected from at least one of carbon (preferably ¹⁴ C), hydrogen (preferably ³ H), sulfur (preferably ³⁵ S), or iodine (preferably ¹²⁵ I). Such radiolabeled probes are conveniently synthesized by radioisotope suppliers specializing in custom synthesis of radiolabeled probe compounds. Such suppliers include Amersham Corporation, Arlington Heights, Illinois; Cambridge Isotop Laboratories, Inc., Andover, Massachusetts; S, Menlo Park, California; SRI International; Wizard Laboratories, West Sacramento, California; Chemex Laboratories, Chemex Laboratories, Lenexa, Kansas; American Radio Label, Chemical Radio, St. Louis, MO (American Rad iolabeled Chemicals, Inc.); and Moravek Biochemicals Inc., Blair, California.

  Tritium-labeled probe compounds can be prepared catalytically by platinum-catalyst exchange in tritiated acetic acid, acid-catalyst exchange in tritiated trifluoroacetic acid, or heterogeneous-catalytic exchange using tritim gas. It is. Such preparation is also conveniently carried out as a custom radiolabel by any of the suppliers described above, using the compounds of the invention as substrates. In addition, certain precursors may optionally be subjected to trithym-halogen exchange using tritium gas, tritium gas reduction of unsaturated bonds, or reduction using sodium borolithiated.

  As described in Non-Patent Document 46, it is possible to perform receptor autoradiography (receptor mapping) in vitro using the radiolabeled compound of the present invention.

[Example]
The following general procedure was used to record analytical data for the following compounds:
Proton NMR spectra were recorded on a Varian VXR or Unity 300 FT-NMR instrument (300 MHz); chemical shifts were deuterochloroform or deuterodimethylsulfoxide as specified below. ) In (ppm) from the tetramethylsilane internal standard. Finnegan MAT 8230 spectrometer or Hewlett Packard 5988A spectrometer (chemical ionization (CI), electrospray (ESI), atmospheric pressure chemical ionization (APCI) or gas chromatography (GC) using NH ₃ as the carrier gas. ))). Melting points are recorded with a MelTemp 3.0 heating block device and are uncorrected. Boiling points are uncorrected. All pH measurements during work-up were performed with indicator paper.

  Reagents were purchased from commercial sources and, if necessary, purified prior to use according to the general procedure outlined in (Non-patent Document 47). Chromatography was performed on silica gel using the following solvent system. For mixed solvent systems, volume ratios are indicated. In other cases, parts and percentages are by weight. Commonly used abbreviations are as follows: DMF (N, N-dimethylformamide), EtOH (ethanol), MeOH (methanol), EtOAc (ethyl acetate), HOAc (acetic acid), DME (1,2-di-). Ethoxyethane) and THE (tetrahydrofuran).

  The following examples are provided to further illustrate the present invention. These examples, which describe the best mode presently contemplated for carrying out the invention, are intended to be illustrative and not limiting.

Preparation A of 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo- [1,3,5] -triazin-4 (3H) -one . 2-methoxy-6-methylpyridine.
To methanol (500 ml) in a flask equipped with a reflux condenser, sodium (31.0 g, 1.35 mol) was added in portions over 30 minutes with stirring. After the addition was complete, the reaction mixture was allowed to cool to ambient temperature. 2-Fluoro-6-methylpyridine (50 g, 450 mmol) was added in portions with stirring. The reaction mixture was then heated to reflux temperature and stirred for 48 hours. The mixture was then cooled to ambient temperature and the solvent was removed in vacuo to give a yellow oil. The residue was taken up in water (500 ml) and extracted three times with ether (200 ml). The combined organic layers were dried over MgSO ₄ , filtered, and the solvent was removed from the filtrate in vacuo to give a yellow liquid: ¹ H-NMR (CDCl ₃ , 300 MHz): δ 7.44 (dd , 1H, J = 8, 7), 6.71 (d, 1H, J = 7), 6.53 (d, 1H, J = 8), 3.91 (s, 3H), 2.45 (s , 3H).

B. 2-methoxy-6-methylpyridine.
A mixture of 2-hydroxy-6-methylpyridine (6.85 g, 62.8 mmol), silver carbonate (22.5 g, 81.6 mmol), iodomethane (39.1 ml, 628 mmol) and chloroform (200 ml) was added in the dark. Stir at ambient temperature for 40 hours. The reaction mixture was filtered through celite. The collected solid was washed with ether. The combined filtrate was concentrated in vacuo to give a liquid (6.25 g), which was identical to the product from Part A.

C. 6-methoxy-3-bromo-2-methylpyridine.
A mixture of 2-methoxy-6-methylpyridine (17.0 g, 138 mmol) and a solution of disodium hydrogen phosphate (0.15 M in water, 250 ml) was stirred at room temperature. Bromine (7.1 ml, 138 mmol) was added dropwise over 15 minutes in the addition funnel. The reaction mixture was then stirred at room temperature for 4 hours. The clear colorless solution was diluted with water (500 ml) and diluted three times with dichloromethane (200 ml). The combined organic layers were dried over MgSO ₄ , filtered, and the solvent was removed from the filtrate in vacuo to give a yellow liquid. Flash chromatography on silica gel (EtOAc: Hexane :: 1: 20) removed the solvent from the desired combined fractions to give a clear colorless liquid (15.4 g). : ¹ H-NMR (CDCl ₃ , 300 MHz): δ 7.60 (d, 1H, J = 8), 6.46 (d, 1H, J = 8), 3.89 (s, 3H), 2. 54 (s, 3H).

D. 6-methoxy-2-methylpyridine-3-boronic acid.
Under nitrogen atmosphere, a solution of 6-methoxy-3-bromo-2-methylpyridine (59.8 g, 296 mmol) in dry THE (429 ml) was cooled to about −78 ° C. with stirring. A solution containing n-butyllithium (2.5 M, 130.4 ml, 326 mmol) in hexane was added dropwise over 30 minutes. The reaction mixture was stirred at about −78 ° C. for 3 hours. A solution of tri-isopropyl borate (102.7 ml, 445 mmol) in dry THE (100 ml) was added dropwise over 30 minutes. The reaction mixture was heated to ambient temperature with stirring for 16 hours. To this reaction mixture was added acetic acid (37.35 g, 622 mmol) followed by water (110 ml) with stirring. After 2 hours, the layers were separated and the organic layer was concentrated in vacuo. The residue was taken up in 2-propanol (750 ml) and the solvent was removed on a rotary evaporator (bath temperature about 50 ° C.). The residue was triturated with ether. The product was collected by filtration and dried in vacuo (48.4 g): melting point> 200 ° C .; ¹ H-NMR (CD ₃ OH, 300 MHz): δ 7.83 (d, 1H, J = 8) ), 6.56 (d, 1H, J = 8), 3.85 (s, 3H), 2.44 (s, 3H); GC-MS: 168 (M + + H).

E. 2-Methyl-3- (5-methylisoxazol-4-yl) -6-methoxypyridine.
4-iodo-5-methylisoxazole (18.2 g, 87 mmol), 6-methoxy-2-methylpyridine-3-boronic acid (14.6 g, 87 mmol), sodium bicarbonate (22.0 g, 262 mmol), A mixture of water (150 ml) and DME (150 ml) was degassed three times by applying vacuum with stirring and then introducing a nitrogen atmosphere. [1,1-Bis (diphenylphosphino) ferrocene] -dichloropalladium (II) (2.14 g, 2.6 mmol) was added in one portion. The reaction mixture was degassed as before. The reaction mixture was then stirred at 80 ° C. for 4 hours and then cooled to ambient temperature. Extracted with EtOAc three times and the combined organic layers were dried over MgSO ₄ , filtered and the solvent removed in vacuo to give an oil. Flash chromatography (EtOAc: Hexane :: 1: 9) and removal of the solvent from the desired fractions in vacuo gave the product (7.15 g): ¹ H-NMR (CDCl ₃ , 300 MHz): δ 8.16 (s, 1H), 7.33 (d, 1H, J = 8), 6.63 (d, 1H, J = 8), 3.95 (s, 3H), 2.35 (s , 6H); APCI ⁺ -MS: 205 (M ⁺ + H).

F. 1-cyano-1- (2-methyl-6-methoxypyrid-3-yl) propan-2-one, sodium salt.
Sodium methoxide (25% w / w, 13 ml, 70 mmol), 2-methyl-3- (5-methylisoxazol-4-yl) -6-methoxypyridine (7.15 g, 35 mmol) and methanol (50 ml) Was stirred at room temperature for 16 hours. The solvent was removed in vacuo to give a yellow oil. Trituration with ether, filtration and drying in vacuo gave the crude product as a white solid (9.3 g).

G. 5-Amino-4- (2-methyl-6-methoxypyrid-3-yl) -3-methylpyrazole.
1-cyano-1- (2-methyl-6-methoxypyrid-3-yl) propan-2-one, sodium salt (9.3 g), hydrazine-hydrate (6 ml, 123.3 mmol) and glacial acetic acid (150 ml) ) Was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuo. The residue was dissolved in 1N HCl and the resulting solution was extracted twice with EtOAc. To this aqueous layer was added 1N NaOH solution until pH = 12. The resulting half solution was extracted three times with ethyl acetate. The combined organic layers were dried over MgSO ₄ and filtered. The solvent was removed in vacuo to give a viscous oil (5.8 g): ¹ H-NMR (CDCl ₃ , 300 MHz): 7.37 (d, 2H, J = 8), 6.62 (d, 2H, J = 8), 3.95 (s, 3H), 2.36 (s, 3H), 2.08 (s, 3H); APCI + -MS: 219 (M + + H); 260 (M + + CH ₃ CN).

H. 5-Acetamidino-4- (2-methyl-6-methoxypyrid-3-yl) -3-methylpyrazole, acetate.
To a rapidly stirred mixture of potassium carbonate (6.95 g, 50.0 mol), dichloromethane (60 ml) and water (150 ml), ethylacetamidate hydrochloride (6.46 g, 52.2 mmol) was added rapidly. . The layers were separated and the aqueous layer was extracted with dichloromethane (2 × 60 ml). The combined organic layers were dried over MgSO ₄ and filtered. The solvent was removed by simple distillation and the container residue (clear light yellow liquid) was used without further purification.

A stirred mixture of 5-amino-4- (2-methyl-6-methoxypyrid-3-yl) -3-methylpyrazole (3.8 g, 17.4 mmol), ethyl acetamidate free base and dichloromethane (100 ml). To this was added glacial acetic acid (1.0 ml, 17.4 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours; at the end, it was concentrated in vacuo. The residue was triturated with ether and the product was filtered and washed with copious amounts of ether. The white solid was dried in vacuo (5.4 g): ¹ H-NMR (CD ₃ OH, 300 MHz): 7.43 (d, 2H, J = 8), 6.69 (d, 2H, = 8), 4.9 (br s, 2H), 3.93 (s, 3H), 2.31 (s, 3H), 2.24 (s, 3H), 2.13 (s, 3H), 1 .88 (s, 3H); APCI ⁺ -MS: 260 (M ⁺ + H).

I. 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo- [1,3,5] -triazin-4 (3H) -one.
Sodium pellet (3.9 g, 169 mmol) was added to ethanol (200 ml) with vigorous stirring. After all the sodium had reacted, 5-acetamidino-4- (2-methyl-6-methoxypyrid-3-yl) -3-methylpyrazole, acetate (5.4 g, 16.9 mmol) and diethyl carbonate (16. 4 ml, 135.3 mmol) was added. The resulting reaction mixture was heated to reflux temperature and stirred for 18 hours. The mixture was cooled to room temperature and the solvent was removed in vacuo. This residue was dissolved in water and 1N HCl solution was added slowly until pH˜6. The aqueous layer was extracted 3 times with EtOAc; the combined organic layers were dried over MgSO ₄ and filtered. The solvent was removed in vacuo to give a solid. Trituration with ether, filtration and drying in vacuo gave a white solid (3.9 g): ¹ H-NMR (CD ₃ OH, 300 MHz): 7.49 (d, 2H, J = 8). ), 6.69 (d, 2H, J = 8), 3.93 (s, 3H), 2.35 (s, 3H), 2.28 (s, 3H), 2.24 (s, 3H) ^{; APCI + -MS: 286 (M} + + H).

4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3,5-triazine Preparation.
A. 4-Chloro-2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolotriazine.
2,7-Dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3,5-triazin-4-one (Example 1, 3.9 g) , 13.7 mmol), di-isopropyl-ethylamine (9.5 ml, 54.7 mmol), phosphorus oxychloride (5.1 ml, 54.7 mmol) and toluene (75 ml) were stirred at reflux temperature for 4 hours. Volatiles were removed in vacuo. The residue was loaded onto a silica gel pad on celite and eluted with a 1: 1 mixture of EtOAc and hexane. The solvent was removed from the filtrate in vacuo to give an oil.

B. 4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3,5-triazine .
4-chloro-2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolotriazine, (R) -2-butylamine (2.0 ml, 20 0.5 mmol), di-isopropyl-ethylamine (9.5 ml, 54.7 mmol) and dry THF (25 ml) were stirred at ambient temperature for 18 hours. The solvent was removed in vacuo. Column chromatography of the residue (first using EtOAc: hexane :: 1: 2, then EtOAc: hexane :: 1: 4) gave the product. The solvent was removed in vacuo to give a white solid (2.3 g): melting point = 18.3 ° C .; ¹ H-NMR (CDCl ₃ , 300 MHz): δ 7.41 (d, 1H, J = 8) 6.63 (d, 1H, J = 8), 6.25 (brd, 1H, J = 9), 4.35-4.30 (m, 1H), 3.95 (s, 3H), 2.49 (s, 3H), 2.35 (s, 3H), 2.30 (s, 3H), 1.76-1.66 (m, 2H), 1.34 (d, 3H, J = 7), 1.02 (t, 3H, J = 7); ¹³ C-NMR (CDCl ₃ , 100.52 MHz): δ 163.8, 163.0, 155.7, 153.7, 147.8 , 146.6, 141.6, 118.5, 107.4, 106.6, 53.3, 48.2, 29.7, 26.1, 22.9, 20.4, 13.1. , 10.3; IR (neat, KBr, cm ⁻¹ ): 3380 (m), 3371 (m), 2968 (m), 2928 (m), 2872 (w), 1621 (s), 1588 (S), 1544 (s), 1489 (s), 1460 (s), 1425 (s), 1413 (s), 1364 (s), 1346 (m), 1304 (s), 1275 (s), 1247 (s), 1198 (m) , 1152 (m), 1134 (m), 1112 (m), 1034 (s), 1003 (m); ESI (+) - HRMS: calculated for _{C 18 H} 24 _{N 6} O Value: 341.2089; Found: 341.2093 (M ^< + ^> + H). _Analysis, C _{18 H} 24 _{N 6} O Calculated: C, 63.51, H, 7.12 , N, 24.69; Found: C, 63.67, H, 7.00 , N, 24. 49.

Rat CRF receptor binding assay to assess useful biological activity.
Receptor binding affinity for rat cortical receptors was assayed according to published methods (Non-Patent Document 48).

Inhibition curves of [ ¹²⁵ I-Tyr ⁶ ] -o-CRF binding to cell membranes at various dilutions of the test drug, the iterative curve fitting program LIGAND [49], provides Ki values for inhibition, Used to assess biological activity].

Inhibition of CRF-stimulated adenylate cyclase activity
Inhibition of CRF-stimulated adenylate cyclase activity can be performed as described in (Non-patent Document 50). Briefly, the assay consists of 100 mM Tris-HCl (pH 7.4 at 37 ° C.), 10 mM MgCl ₂ , 0.4 mM EGTA, 0.1% BSA, 1 mM isobutylmethylxanthine (IBMX), 250 units / ml phosphocreatine. Kinase, 5 mM creatine phosphate, 100 mM guanosine 5 ′ triphosphate, 100 nM oCRF, antagonist peptides (concentration range 10 ⁻⁹ to 10 ⁻⁶ m) and raw wet weight tissue 0.8 mg (approximately 40-60 mg protein) Carry out for 10 minutes at 37 ° C. in 200 ml of buffer. The reaction is initiated by adding 1 mM ATP / [ ³² P] ATP (approximately 2-4 mCi / tube) and terminated by adding 100 ml of 50 mM Tris-HCl, 45 mM ATP and 2% sodium dodecyl sulfate. To monitor cAMP recovery, 1 μl (approximately 40,000 dpm) of [ ³ H] cAMP is added to each tube prior to separation. Dowex (Dowex) and aluminum column, by sequential ^elution, to separate the ^[32 P] cAMP from [32 P] ATP.

In Vivo Biological Assays Any one of the biological assays available and recognized within the art can be used to assess the in vivo activity of the compounds of the present invention. Examples of these tests include an auditory startle assay, a stair climbing test, and a chronic administration assay. These and other models useful for testing compounds of the present invention are outlined in (51).

  Compounds can be tested in all species of rodents or small mammals.

  The compounds of the present invention are useful for the treatment of imbalances associated with abnormal levels of corticotropin releasing factor in patients suffering from depression, affective disorders, and / or anxiety.

  The compounds of the present invention can be administered to treat these abnormalities by a method of contacting the active agent with a site of drug action within the mammal. The compounds can be administered in a conventional manner that can be utilized in conjunction with pharmaceuticals as either a single therapeutic agent or a combination of treatments. While the compounds of the invention can be administered alone, they are generally administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

  Dosage depends on known factors such as the use and pharmacodynamic properties of the particular drug, and its method and route of administration; age, weight, and health of the recipient; nature and extent of symptoms; type of current treatment; Depending on the frequency of; and the desired effect. When used in the treatment of the above diseases or conditions, the compound of the present invention may be orally administered daily at a dosage of the active ingredient of 0.002 to 200 mg / kg body weight. In general, a single dose of 0.01-10 mg / kg in divided doses, 1 to 4 times a day, or a sustained release formulation will be effective in obtaining the desired pharmacological action.

  Dosage forms (composition) suitable for administration contain from about 1 mg to about 100 mg of active ingredient per unit. The active ingredient is usually present in these pharmaceutical compositions in an amount of about 0.5 to 95% by weight, based on the total weight of the composition.

  Active ingredients that can be administered orally are solid dosage forms such as capsules, tablets and powders; or liquid formulations such as elixirs, syrups and / or suspensions. The compounds of the present invention can also be administered parenterally in sterile liquid formulations.

  Gelatin capsules can be used to contain the active ingredient and a suitable carrier such as, but not limited to, lactose, starch, magnesium stearate, stearic acid, or cellulose derivatives. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release over time. Compressed tablets can be sugar coated or film coated to mask unpleasant taste, or to protect the active ingredient from the atmosphere or to allow selective disintegration of the tablets in the gastrointestinal tract used.

  Liquid dosage forms for oral administration may contain coloring or flavoring to increase patient acceptance.

  In general, water, pharmaceutically acceptable oils, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycol are suitable carriers for parenteral solutions. A solution for parenteral administration preferably contains a water-soluble salt of the active ingredient, an appropriate stabilizer, and if necessary, a butter-like substance. Antioxidants, such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or in combination, are suitable stabilizers. Citric acid and its salts and EDTA are also used. In addition, parenteral solutions may contain preservatives, such as benzalkonium chloride, methyl paraben, propyl paraben, and chlorobutanol.

  Suitable pharmaceutical carriers are described in (52), a standard reference in the field.

  The pharmaceutical dosage forms useful for administering the compounds of the invention are described below by way of example.

Capsules A number of unit capsules are prepared by filling each standard two-piece hard gelatin capsule with 100 mg powdered active ingredient, 150 mg lactose, 50 mg cellulose, and 6 mg magnesium stearate.

Soft gelatin capsule A mixture containing the active ingredient in easily digestible oil such as soybean oil, cottonseed oil, or olive oil is prepared and injected into gelatin with a positive displacement pump to form a soft gelatin capsule containing 100 mg of the active ingredient To do. The capsule was washed and dried.

Tablets A number of tablets were prepared by the conventional procedure where the dosage unit was 100 mg active ingredient, 0.2 mg colloidal silicon dioxide, 5 mg magnesium stearate, 275 mg microcrystalline cellulose, 11 mg starch, and 98.8 mg lactose. Is done. Appropriate coatings can be applied to enhance palatability or delayed adsorption.

  The compounds of the invention can also be used as reagents or standards in biochemical studies of neurological function, dysfunction, and disease.

  Although the invention has been described and illustrated with respect to certain preferred embodiments, other embodiments will be apparent to those skilled in the art. Accordingly, the invention is not limited to the specific embodiments described and illustrated, but may be modified or changed without departing from the spirit of the invention, the entirety of which is set forth in the appended claims. Indicated by.

Claims (4)

Formula (I):

Or a pharmaceutically acceptable salt thereof . The compound is 4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3, The compound according to claim 1, which is 5-triazine, or a pharmaceutically acceptable salt thereof . 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof , wherein the compound is substantially free of its (S) stereoisomer. The compound is 4-((R) -2-butylamino) 2,7-dimethyl-8- (2-methyl-6-methoxypyrid-3-yl) [1,5-a] -pyrazolo-1,3. The compound of claim 1, which is 1,5-triazine.